Protein Isolation
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Descripción
Experiment Principles of Protein Expression, Isolation and Purification Expression of gene clone in cells is of great significance in both theoretical research and experimental applications. We can explore and study...
mostra másExpression of gene clone in cells is of great significance in both theoretical research and experimental applications. We can explore and study gene function and expression mechanism expression with gene expression. What’s more, the encoded protein by cloned gene expression can be used in researching the structure and function.
E.coli (Escherichia coli ), whose exogenous expressing productivity is much higher than that of other gene expression systems, is currently the most widely used protein expression system. The number of target protein expressed may even exceed 80% of the total protein in bacteria. In this experiment, the plasmid carrying target protein overexpresses recombinant chloramphenicol acyltransferase protein with 6 consecutive histidine residues in E. coli BL21 under the induction of IPTG in 37 ℃. The protein can be purified with chromatographic media formed by immobilizing nickel ions (Ni2 +) with covalent coupling nitrilotriacetic acid (NTA), which is in fact Metal Chelate Affinity Chromatography (MCAC). The purity of the protein can be analyzed by polyacrylamide gel electrophoresis. https://www.creative-biolabs.com/blog/index.php/protein-expression-isolation-and-purification/
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